JP5133411B2 - Collection / extraction vessel for biological material in forensic samples - Google Patents

Description

  The present invention relates to a collection / extraction container for collecting solid forensic samples and / or for extracting biological material from those forensic samples. The container has an upper opening for inserting a forensic sample. The upper opening is surrounded by a wall that surrounds an internal chamber that is essentially a vertical wall and whose depth is limited by the bottom surface. The walls and bottom of the container are impermeable to fluid. The container also comprises a basket and an essentially horizontal intermediate floor for storing solid forensic samples during washing or digestion / lysis, extraction. The intermediate floor is permeable to fluid and divides the internal chamber into an upper sample space and a lower fluid space.

  The collection of samples containing nucleic acids to determine the genetic code is currently increasing in importance. Regarding the fight against crime, the collection of genetic samples in the sense of “gene fingerprint” has become increasingly important in two respects. First, the genetic code should be obtained from a criminal who has already been arrested, and second, the data obtained should be compared with unknown evidence found at the crime scene, for example. is there. In the first case, a fresh and obvious sample is used to build the data bank, which is accessible in the second case. However, in the second case, the collected samples are often incomplete, dirty and damaged, complicating forensic work. Furthermore, individual countries have already begun to collect biological samples to prophylactically obtain the genetic code of all individuals entering the country or of all residents of that country.

  The “forensic” attribute refers to anything that has legal or criminal characteristics. The term is therefore not limited to the field of criminal law (eg forensic medicine) but rather includes any professional activity within any legal process. Samples suitable for court are also proteins (eg prions that cause Creutzfeldt-Jakob syndrome, bovine spongiform encephalopathy, or BSE, respectively), viruses, bacteria, and other microorganisms, human or animal body fluids (eg blood, saliva, Feces, semen, and urine), and single cells (such as oral mucosal cells and hair follicles).

  Methods for isolating and analyzing human deoxyribonucleic acid (DNA) ribonucleic acid (RNA) are known from the prior art (eg, Molecular Diagnostics, Isolation and Analysis of Human Genomic DNA, 1998, Promega Notes No. 68, page 20). It has been. These methods comprise the PCR (PCR = polymerase chain reaction) method, which is itself well-known for increasing sample production and thus analytical sensitivity.

  A container for collecting samples and manually preparing the collected samples for analysis or PCR reaction is known from WO 2004/105949 A1. Containers for performing collection and subsequent on-site PCR reactions are known from US 2004/0214200 A1. However, the known methods are very complex and the container appears unsuitable for processing samples automatically and / or robotically.

  Automated nucleic acid separation is known, for example, from US 5,863,801. However, the device described in this patent is designed to be used as a stand-alone device and is based on TECAN's automated liquid handling system (TECAN Schweiz AG, Seestrasse 103, CH-8708 Mannedorf, Switzerland). Does not work with existing automated equipment commonly used in forensics and research laboratories. In order to validate their processing, the means required in such laboratories are provided, and have consumables that mimic those validated manual processes and can be used with existing equipment. It is an important value.

  An apparatus for culturing, centrifuging, and separating DNA samples on a solid support is known as the Slicprep ™ 96 apparatus from PROMEGA (2800 Woods Hollow Road, Madison, WI 53711 USA). This device is made from polypropylene and is based on 96 deep well microplates, 96 spin basket units, and a collar to raise the basket. In the culture method, a solid support or forensic sample (eg, a dry oral swab) is placed in a basket that is fully inserted into a deep well plate. Degradation buffer or lysis buffer is added to cover the sample and the sealed unit is incubated at the desired temperature. The basket is then raised and the collar is inserted so that centrifugation in a rocking plate rotor removes the extract, i.e. the DNA-containing solution, from the solid support or forensic sample. After removing 96 rotating basket units and collars, the deep well plate can be placed on a workstation for DNA purification. This device was designed to make DNA extraction from forensic samples more automatically compatible, but still requires manual procedural steps to add color and then remove the color and basket unit.

  Nucleic acid separations are performed from FITZCO (4300 Shoreline Drive, Spring Park, MN 55384, USA) which are carried out as described above and require significant manual steps during processing. The FITZCO Spin-eze (TM) device is a single tube device consisting of a basket unit and an Eppendorf-type receiving tube. In the first extraction step, a solid support (ie, a cotton swab) containing DNA is provided in the basket, after which it is inserted into an Eppendorf-type receiving tube. Degradation or lysis buffer is added to cover the forensic sample and the sealed unit is incubated and then centrifuged to flow the solution containing DNA from the basket to the tube. The basket is then manually removed and the tube can be placed on a workstation to purify the DNA.

  Another system known from TECAN consists of two tubes, which can be attached end-to-end with a “filter” placed between them for processing. The sample tube contains a swab or other DNA-containing forensic material. For initial processing, buffer is added to the sample tube and the receiving tube is attached upside down to the top of the sample tube. A rack having such an array of “double tubes” is moved to a “reversal” device that, during subsequent centrifugation, liquid is now passed from the upper sample tube through the filter into the lower receiving tube. Rotate the two connected tubes so that they can flow. The tube can then be separated. This system also requires manual operation and requires a special device to invert the tube.

WO 2004/105949 A1 US 2004/0214200 A1 US Pat. No. 5,863,801

  The object of the present invention proposes an alternative device and / or an alternative method that can process bound molecules such as nucleic acids and that can be completely automatically removed from a solid support such as an oral swab. That is.

This object is achieved according to the first aspect by a collection / extraction container for collecting solid forensic samples and / or for extracting biological material from forensic samples according to the features of claim 1. . The container has an upper opening for insertion of forensic samples. The top opening is surrounded by an essentially vertical wall that surrounds an internal chamber whose depth is limited by the bottom surface. The walls and bottom are impermeable to fluid. The container comprises a basket for holding solid forensic samples during washing, ie during decomposition / dissolution and extraction, and an essentially horizontal intermediate floor, which is permeable to the fluid. The internal chamber is divided into an upper sample space and a lower fluid space. The container comprises an essentially vertical channel, which achieves allowing the insertion of a pipetting needle through the sample space and into the fluid space, and the channel is open at the top of the container With a top orifice located near the bottom orifice and through the intermediate floor at the bottom orifice. Here, the container also includes a partition wall that at least partially surrounds the flow path and separates the latter from the upper sample space.

In the container according to the present invention, the basket is installed in the collection / extraction container coaxially with the central axis of the collection / extraction container, and the basically vertical flow path is basically the collection / extraction container. The flow path is disposed off-center with respect to the central axis so as to be partially formed by a vertical wall and a partition wall.

This object is achieved according to a second aspect according to the features of claim 17 in which a method for extracting biological material from a forensic sample is proposed. The method according to the present invention comprises the following steps. That is,
(A) providing at least one container 1 with an accessible upper opening 3, an upper sample space 9 with a basket 7, a fluid space 10 and a flow path 11;
(B) storing the solid forensic sample 2 through the upper opening 3 into the basket 7 in the upper sample space 9 of at least one container 1;
(C) adding decomposition / lysis buffer into the basket 7 so that the lower fluid space 10 of the container 1 is filled and the upper sample space 9 is also sufficiently filled to cover the forensic sample 2;
(D) sealing the whole apparatus 1 and then allowing culturing to allow an extraction step to take place;
(E) After culturing, the device 1 is opened or opened, and the degradation buffer is removed by accessing the bottom surface of the lower fluid space 10 through the basically vertical channel 11;
(F) Place the device 1 in a centrifuge and rotate it so that any buffer remaining in the solid forensic sample 2 collects down to the bottom of the internal chamber 5;
(G) removing the remaining decomposition buffer collected at the bottom of the lower fluid space 10 by centrifugation through the basically vertical flow path 11;
Here, the solid forensic sample 2 is held in the basket 7 in the sample space 9 of at least one container 1 between all steps (c) to (g).

  Preferably, the removed volume is combined with the previously removed volume. The combined sample is ready for nucleic acid purification using a commercially available kit.

  Additional preferred features according to the invention result from the dependent claims.

The advantages of the present invention include the following points. That is,
-For example, the same containers used for the extraction of biological material from the surface of solid forensic samples can also be used for the collection of such forensic samples in the field.
The container provides a port or flow path that allows the pipette tip to access the bottom of the container without the need for removal of the basket containing the solid forensic sample.
-The containers are arranged, for example in a 6x8 tube array, so that it is not necessary to assemble and disassemble the parts during processing and can be fully automated. Therefore, ordinary automatic devices such as centrifuges, robot arms, PCR thermostat tables, etc. can be used.
-There is no need for special equipment to invert the tube or rack containing the tube.
-Labeling the container with an ID tag provides a clear identification of a particular forensic sample and biological material extracted from this solid forensic sample.

FIG. 3 shows a vertical bottom section through two different racks with four different embodiments of containers according to the invention, with a flat bottom container with a basket fitted in friction. FIG. 4 shows a vertical partial cross section through two different racks with four different embodiments of a container according to the invention, shaped like a PCR tube, and a two-part container. FIG. 2 is a horizontal partial cross-section of the two racks of FIG. 1, showing a collection rack having an insertion basket that fits frictionally. FIG. 2 is a partial cross-section in the horizontal direction of the two racks of FIG. 1 and shows an extraction rack having containers held in place by holding means. Figure 2 is a vertical partial cross section through a rack having two different embodiments of containers according to the present invention.

The invention will be explained in more detail on the basis of exemplary embodiments and schematic drawings. These do not limit the scope of rights of the present invention.
FIG. 1 shows a vertical partial section through two different racks having four different embodiments of a container 1 according to the invention. FIG. 1A shows a flat bottom container with a basket fitted in friction. Both containers shown are collection / extraction containers 1 for collecting solid forensic samples 2 and / or for extracting biological material from these forensic samples 2. The solid forensic sample 2 is generally selected from the following group. The group includes gauze pads or swabs; findings; filter papers or WHATMAN FTA papers (Whatman plc., 27 Great West Road, Brentford, Middlesex, TW8 9BW, UK, respectively); solids, nucleic acids or preparative gels and Sheet-like material for absorbing other biomolecules such as cellulose membranes; and textile pieces. In most cases, the biological material sample to be extracted is selected from the group comprising body fluids, cells, DNA, RNA, proteins, microorganisms, and viruses. However, it is preferred to collect nucleic acid material such as DNA and / or RNA with an oral swab and extract the nucleic acid material from the oral swab in the container 1 of the invention.

  Each container 1 has an upper opening 3 for inserting a solid forensic sample 2. The top opening 3 is surrounded by an essentially vertical wall 4 that surrounds an internal chamber 5 that is limited in depth by a bottom surface 6. The wall 4 and the bottom surface 6 are impermeable to fluid. Each container 1 comprises a basket 7 and an essentially horizontal intermediate floor 8 for holding a solid forensic sample 2 during washing, ie during decomposition / dissolution, extraction. The intermediate floor 8 is permeable to fluid and divides the internal chamber 5 into an upper sample space 9 and a lower fluid space 10.

  In the context of the present invention, “fluid” is to be understood as a liquid, a gas or a liquid / gas mixture. “Solid” is interpreted as a hard or soft substance other than a fluid. A “sample” is preferably a solid forensic sample, but it can be another sample containing biological material. The “findings” can be artifacts or casework samples, such as those found in crime scenes, or any other unspecified solid material including extractable biological material.

  The container 1 according to the invention comprises a basically vertical channel 11. The channel 11 accomplishes allowing the pipetting needle 12 to be inserted through the sample space 9 and into the fluid space 10. This is achieved by the passage 11 having a top orifice 13 located near the top opening 3 of the container 1 and the passage 11 passing through the intermediate floor 8 at the bottom orifice 14. The container 1 also includes a partition wall 15 that at least partially surrounds the flow path 11 and separates the latter from the upper sample space 9.

  Preferably, the wall 4 of the container 1 is essentially cylindrical at least in the region of the upper sample space 9. In order to leave as much space as possible for the solid forensic sample 2 that can be introduced into the upper sample space 9 and can be maintained by the basket 7, the channel 11 is arranged off-center and is separated by the wall 4 and the partition wall 15. It is preferred that it is partially formed. It is particularly preferred that the basket 7 of the container 1 is sized to accommodate an oral swab.

  Preferably, the channel 11 is arranged off the center, so that a vertical axis 40 is formed which is essentially off center. Thus, the bottom surface 6 and / or the lower section 17 of the wall 4 is preferably finished to form a fluid collection region 21. The fluid collection region 21 is at the lower level, located in the rest of the bottom surface 6 of the container 1 and is essentially on a vertical axis 40 off center.

  As shown in FIG. 1A, the basket 7 may be finished as a single injection molded article of polymeric material that is inserted into the internal chamber 5 of the container 1. Also, as shown in FIG. 1B (see its right side), the basket 7 is finished as part of the upper section 16 of the wall 4 and together with the upper section 16 of the wall 4 as a single injection molded article of polymeric material. It is formed. The lower section 17 of the wall 4 advantageously comprises a socket 19 at the upper end 18 into which the lower end 20 of the upper section 16 of the wall 4 can be hermetically inserted. Further, as shown in FIG. 1B (see the left side thereof), the container 1 may be shaped like a PCR tube at least in the region of the lower fluid space 10.

  The bottom surface 6 of the container 1 is preferably finished by forming a single injection-molded article of polymer material together with the wall 4 or the lower section 17 of the wall 4. A preferred polymeric material for injection molding of the parts of the container 1 according to the invention is polypropylene. Instead, the bottom surface 6 of the container 1 is finished as a film structure 27 that is glued or welded to the wall 4 or the lower section 17 of the wall 4 (see eg FIG. 1A, left side).

  The bottom surface 6 is preferably provided with a flat region 22 on the outside thereof, and an information tag 23 is placed on or in the flat region 22 (see, for example, FIG. 1B, right side). The information tag 23 is advantageously selected from the group comprising 1-D barcode, 2-D barcode, RFID transponder, and RuBee transceiver. The principles of 1-D barcodes and 2-D barcodes are well known to those skilled in the art and are based on optical scanning of high contrast identification markings. The advantage of such identification is that it is a relatively simple physical principle, however, the scanning device and information tag must be in view.

  RFID transponders are also known per se and operate at high frequency (HF, eg 900 MHz) or ultra high frequency (UHF). They send and receive radio signals. On the other hand, newer RuBee transceivers operate at wavelengths below 450 kHz, transmit and receive signals, and are primarily based on magnetism. Passive RFID transponders can receive approximately 100 (HF) or 150-200 (UHF) messages per second. In contrast, an active RuBee transceiver can only receive approximately 10 messages per second. That is, in either case, visual contact is not necessary. Thus, the type of information tag 23 used is a function of, among other things, the density of data transfer and the presence of visual contacts. In the case of FIG. 1B, the RFID transponder or RuBee transceiver is located on the vertical wall 4 of the container 1 (see left side), while the 2-D barcode is located in the flat area 22 of the bottom surface 6 (right side). See). Any arbitrary combination of barcodes and RFID transponders or RuBee transceivers is conceivable, such as simple 1-D barcodes are used to identify forensic samples, especially during field collection, for example. This is the case when RFID transponders or RuBee transceivers are used during the automatic extraction of relevant biological materials at a high-throughput workstation in a forensic laboratory.

  The container 1 preferably includes a closing member 24 for the upper opening 3. Preferably, depending on the portion of use, the closure member 24 is selected from the group comprising a plug 25, a cover 26 and a film 27. As an example, a collection set 28 is shown in FIG. 1A. This collection set 28 has at least one collection / extraction container 1 according to the invention. The collection set 28 comprises a collection rack 29 having a compartment 30 for receiving containers.

  Preferably, the compartment 30 of the collection rack 29 is open at the top and bottom. As a result, the container 1 can be inserted into the compartment 30 from above or below and can be discharged downward or upward from this compartment. It is particularly preferred that the collection rack 29 has microplate standard dimensions. Standards regarding the dimensions of the microplate are officially published by ANSI (American Standards Association). These standards include the Foodprint Dimensions (ANSI / SBS 1-2004), the Height Dimensions (ANSI / SBS 2-2004), Bottom Outside Flange Dimensions (ANSI / SBS 3-2004), the Well Positions (ANSI / SBS 4 -2004) defines microplates. However, strip racks with 6 or 8 compartments in a row can also be selected for collecting forensic samples.

  If the collection set 28 originally includes sample tubes or containers 1 without individual closure members, a film 27 covering the entire collection rack 29 is preferred on the bottom as well as the top (see FIG. 1A, left). . Thus, a completely clean container free of any contamination is provided at the site where forensic samples are collected. As soon as the fresh sample 2 is put into the basket 7 of the container 1, a stopper 25 can be fitted to completely close the upper opening 3 of the container 1 (see FIG. 1A, right). The stopper 25 can include a barcode (not shown here) that identifies the individual forensic sample 2 in the container 1.

  As an alternative, the collection set 28 may originally comprise a sample tube or container 1 with individual closure members 25. In this case, at least the film 27 at the upper end of the collection rack 29 can be eliminated. Further, if the bottom surface of the compartment 30 is not covered with the film 27, the barcode can be supplied on the flat area 22 of the container bottom surface 6 (not shown here). Eventually, when the barcode cannot be used, the container is classified only by the RFID transponder or the RuBee transceiver. Such intelligent labels can also include additional data that is input prior to or during sample collection and all further processing and is readable by a dedicated device.

  In the laboratory, the sample can be processed in a collection rack 29. However, it is preferred to collect containers from different collection racks 29 and place them into one or more extraction racks 33. Such extraction racks 33 are standardized to allow them to be transported by a microplate robot and handled at a microplate handling station in a laboratory workstation. It preferably has the shape and size of a microplate. One extraction rack 33 and at least one container 1 inserted into a compartment 30 of this rack form an extraction set 32.

  A container 1 with a solid forensic sample 2 may be inserted into any compartment 30 of such an extraction rack 33 by a robot (not shown). In order to achieve this, both racks 29, 33 are arranged on top of each other so that both the collection rack 29 and the extraction rack 33 coincide with each other. With either one of the two racks at the top of the other, the robot tool is used to push the container 1 from above into the compartment of the extraction rack 33 from below. This operation is greatly facilitated in that both types of racks 29, 33 have compartments 30 that are open at the top and bottom, so that the container 1 can be inserted into the compartment 30 from the bottom or top. It is possible to take out from the compartment 30 downward or upward.

  Individual processing of each container 1 by a robot is also possible, even if processing of the containers 1 is preferably performed in racks 29,33. That is, the container 1 does not necessarily have to be placed in a rack in order to be processed in an automated manner. Indeed, for example, TECAN devices can move individual tubes using so-called “pick and place arms”. In fact, for casework samples in forensic research, it is rather preferable to process a small number of samples (eg, 1-8) simultaneously and wait for 96 samples. Even with a small number of samples, automation offers advantages such as higher reproducibility, "control of processing and distribution processes" and walk away time.

  As shown in the horizontal partial cross-section of the two racks 29, 33 in FIG. 2, the container 1 is preferably held in the compartment 30 of the racks 29, 33 using the holding means 35 (see FIG. 2B). ). However, the container 1 can be located in the compartment only by a friction fit (see FIG. 2A). The container 1 preferably has an upper flange 36 and a lower wall thickness region 37 which advantageously have the same outer diameter. This diameter is slightly larger than the distance between the two parallel intermediate walls 34 of the collection or extraction racks 29,33. Thus, the insertion of the container slightly deforms the container and the four adjacent intermediate walls 34, and the friction between the upper flange 36 and the lower wall thickness region 37 and the inner surface of the intermediate wall 34 is reduced in the compartment 30. Keep the container inside.

  2A and 2B are partial cross-sections in the horizontal direction of the two racks 29 and 33 as shown in FIG. In the right compartment of FIG. 1A and the left compartment of FIG. 1B, the bottom surface 6 of the inserted container 1 (see FIG. 1A) or the lower section 17 of the wall 4 (see FIG. 1B) defines the fluid collection area 21. Formed and finished. The fluid collection area 21 is the remaining part of the bottom surface 6 at the lower level of the container 1. This fluid collection region 21 is on an axis 40 that is essentially off-center. The relative position of the two containers relative to the inserted basket 7 (see FIG. 1A, left) or the integrated basket 7 (see FIG. 1B, right) is not of interest, whereas the fluid collection area The relative positions of the flow paths 11 of the two containers having 21 are important. In order to define the exact relative position of the respective collection area 21 in the basket 7, the channel 11 and the bottom surface 6 of the container 1, the basket is preferably equipped with an essentially vertical orientation bar 41. . The container includes in the region of the upper sample space 9 an essentially vertical orientation groove 42 shaped to receive the orientation bar 41 of the basket 7. With this orientation bar and groove 41, 42 design, the desired position of the basket 7 defines its orientation about the central axis 43 of the container 1 as well as its height, ie its distance from the bottom surface 6 of the container 1. be able to.

  While deviating from the embodiment of the intermediate floor 8 having a square opening as shown here, without departing from the spirit of the present invention, the intermediate floor is permeable to liquids by any through-opening other than square. It is possible to make it. A material different from the polymer used to mold the basket 7 and / or the container 1 can be used for the intermediate floor 8. Such materials include filter paper and fabrics, natural and / or synthetic fibers, and / or metal wires. However, it is particularly preferred that the entire container and basket are made using only one material so that the container 1 as well as the basket 7 is preferably designed as an inexpensive consumable.

  The desired position of the basket 7 with respect to the distance from the bottom surface 6 of the container 1 can also be defined by the formation of a shoulder 44 on the inner surface of the wall 4. When the inner diameter of the upper sample space 9 is larger than the inner diameter of the lower fluid space 10 of the container 1, a shoulder 44 is formed. Accordingly, the outer diameter of the basket 7 is larger than the inner diameter of the lower fluid space 10, and the basket 7 rests on the shoulder 44 (see FIG. 1A, left).

  FIG. 3 shows a vertical partial cross section of an extraction rack 33 having two different embodiments of the container 1 according to the invention. The cross section and the illustrated direction are indicated by arrows in FIG. 2B. In addition, these containers 1 preferably have an upper flange 36 and a lower thick region 37 having the same outer diameter. The upper flange 36 and the lower wall thickness region 37 create a friction lock with retaining means 35 and are preferably located in the corners of the compartment 30 and extend essentially beyond the overall height of the compartment 30. Thanks to this friction lock, the container can be held firmly in the compartment 30 and not fall off naturally. Furthermore, these holding means 35 can have first and second projections 38, 39, upon insertion into the compartment 30, between which the lower thick region 37 of the container 1 is engaged, As a result, a specific height position of the container 1 in the compartment 30 is defined in advance, and the holding at the defined position is further strengthened. Friction locks with the container 1 inserted can be defined by the exact dimensions of their holding means 35, but can also be defined by their flexibility range.

  The container 1 in the left compartment of FIG. 3 is identical to that of the right compartment of FIG. 1B, but the position and size of the socket 19 at the upper end 18 of the lower section 17 of the wall 4 is different. That is, the position of the socket is lower and the height of the socket 19 is higher. In both cases, however, the intermediate floor 8 is formed like a cobweb-like lattice that is manufactured in one piece with the upper section 16 of the wall 4. The ability to make a seal between two areas of the wall 4 increases with the size of the socket 19.

  The container 1 preferably comprises a closing member 24 for the upper opening 3. For convenience at the site of use, the closure member 24 is selected from the group comprising a plug 25, a cover 26, and a film 27. As an example, a collection set 28 is shown in FIG. 1A. This collection set 28 has at least one collection / extraction container 1 according to the invention. The collection set 28 comprises a collection rack 29 having a compartment 30 for receiving containers.

  Since liquid can be automatically added to the solid sample, a pipetting needle 12 can be introduced into the liquid space 10 through the channel 11 (see FIG. 1B, left), or the dispenser needle can be solid forensic. It can be located above the sample 2 (see FIG. 3). Also, a pipetting needle 12 can be introduced through the channel 11 to automatically remove liquid from the liquid space 10 (see FIG. 1B, left). In all cases, access is required through the upper opening 3 of the container 1. The top opening 3 can be completely closed by a stopper 25 mounted after collecting forensic samples or placed in the container between two preparatory steps at the laboratory workstation (right of FIG. 1B). reference). The upper opening 3 can also be partially closed by a cover 26 that covers only the sample space 9 and leaves the channel 11 open (see FIGS. 1B and 2B, left). If the container 1 comprises an essentially vertical orientation groove 42 shaped to accommodate the orientation bar 41 of the basket 7 in the region of the upper sample space 9, the cover 26 preferably also comprises an orientation bar 41 (FIG. 2B, see left).

All plugs 25 and covers 26 are preferably provided at their upper ends with a gripping tube 31 (see, for example, FIG. 1B), which gripping tubes 31 are used to remove the plugs 25 or cover 26 or to the container 1. It can be gripped by a robot (not shown) for proper attachment. Although deviating from the illustrated embodiment, but without departing from the spirit of the invention, certain knobs (eg, “Eppendorf” tubes, which allow a person to easily remove and open the plug 25 or cover 26, (Not shown) can be used as an alternative solution. The user can manually open the containers 1 before placing them in the automation system.
Again, in order for a single sample to be provided in the basket, in another embodiment, a plug 25 or cover 26 can be included with a septum piercable above the basket 7 and / or the channel 11, The cover 26 does not need to be removed, and the sample space 9 and / or the channel 11 can be accessed by penetrating the plug 25 or the partition wall of the cover 26 (not shown).

The present invention also provides a method for extracting biological material from a solid forensic sample 2. This method according to the invention comprises: That is,
(A) providing at least one container 1 according to the invention having an accessible upper opening 3, an upper sample space 9 with a basket 7, a fluid space 10 and a flow path 11;

  (B) A solid forensic sample, typically a swab head, cloth, WHATMAN FTA paper, or other material containing nucleic acids, through the upper opening 3 into a basket 7 in the upper sample space 9 of at least one container 1. Place 2;

  (C) sufficient degradation / lysis buffer {general proteinase) into the basket 7 so that the lower fluid space 10 of the container 1 is filled and the upper sample space 9 is also filled enough to cover the forensic sample 2 K solution; commercial examples include PROMEGA DNA IQ Lysis Buffer, QIAGEN Proteinase K (QIAGEN AG, Garstligweg 8, CH-8634 Hombrechtikon, Switzerland) and INVITROGEN ChargeSwitch <(R)> Lysis Buffer L13 (Invitrogen Corp., 1600 Faraday Avenue , Carlsbad, CA 92008, USA}.

  The actual amount will depend on the size of the device. That is, it will depend on different sized devices for different size solid forensic samples 2. In one embodiment, container 1 will be sized similar to a typical 2 ml microcentrifuge tube with a basket that will contain a volume of about 1.25 ml. Such an embodiment would require adding about 1 to 1.25 ml of buffer in this step.

  (D) The whole apparatus is sealed with the film seal 27, the closing member 24, the stopper 25, or other means, and the culture is enabled so as to enable the extraction process. Culture times and temperatures recommended by the manufacturer of the lysis buffer will be used (temperatures of 56 ° C, 70 ° C, and 95 ° C are often used for a time range of up to 1 hour or more).

  (E) After culturing, the apparatus is opened or opened, and the decomposition buffer solution is taken out by accessing the bottom surface of the lower fluid space 10 through the basically vertical channel 11. The amount removed can be the total recoverable amount, but must be greater than or equal to the volume of the upper sample space 9 plus the amount of liquid absorbed by the solid forensic sample 2. This recovered amount of buffer should be saved in another user-selected vial / tube / etc. For later processing.

  (F) Place the device 1 in a centrifuge and rotate it so that any buffer remaining in the solid forensic sample is collected at the bottom of the internal chamber 5, preferably in the fluid collection area 21. The centrifuge should be set at a speed and time that maximizes fluid recovery from the solid forensic sample.

  (G) The remaining amount of the decomposition buffer collected in the bottom of the lower fluid space 10, preferably in the fluid collection region 21, is removed by centrifugation through an access through the basically vertical flow path 11. The removed quantity is integrated with the previously removed quantity. This sample is ready for nucleic acid purification using, for example, commercial nucleic acid purification kits commercially available from PROMEGA, INVITROGEN, QIAGEN, and other companies.

  In this device 1, the volume of lysis buffer containing nucleic acid “washed out” the solid forensic sample 2 is collected through the “access channel” 11 so that the liquid can be removed and the solid sample 2 remains behind. Thus, according to the invention, the lysis buffer can be removed by the pipetting needle without having to remove the basket 7 containing the solid forensic sample. In particular, this feature greatly facilitates the use of the container 1 in fully automated workstations such as TECAN type workstations including devices for sealing / opening and centrifuging (typical options in these instruments). .

In using the container according to the invention, it is optional to perform the following A or B. That is,
A Extracted liquid and biological material extracted by the pipetting needle 12 inserted into the flow path 11 and passing through the sample space 9 to reach the fluid space 10 are taken out, and amplification and / or analysis of the extracted biological material is performed in the container 1. Performing outside or B Amplifying the extracted biological material in the fluid space 10 of the container 1.

  In any case, however, the culturing step is preferably performed in step (d) and the container 1 is subjected to a centrifuge after performing one of the steps (e).

  The biological material to be extracted from the solid forensic sample can be anything such as biological material. However, it is preferred to select this biological material from the group comprising body fluids, cells, DNA, RNA, proteins, microorganisms, and viruses. In particular, biological materials in the form of nucleic acids or nucleotides are preferred.

  In order to be able to carry out the method according to the invention on a workstation, such a workstation is subjected to chemical analysis, for example with chemicals and biochemical compounds and / or substances on a regular basis for potential pharmaceutical effects. It is preferably equipped with a corresponding control system as is known per se in the field of liquids handled in pharmaceutical research.

  In such known automated systems, the containers are transported and manipulated by special equipment called a “workstation”. These workstations can be manually operated individually or connected with automated systems. With the automated system, the user does not have to perform or have to carry out all the methods of the individual processes. Another common factor that couples such known systems is the fact that samples are often processed in standardized microplates. Such microplates can be obtained in any configuration, but in general 96 sample containers or "", arranged in a regular 8x12 raster (according to ANSI standards) with a 9mm spacing between the centers. Well ". A microplate having wells of multiples of this number, or only a part thereof, is also used. Different workstations may be connected to one or more robots to carry the microplate. One or more robots, preferably moving according to a Cartesian system, may be used at the top of the workbench. These robots can carry plates or other sample containers and also fluids. A central control system or computer monitors and controls those known systems, the notable advantage being the complete automation of the work process. As a result, such a system can be operated continuously for hours or days without the need for any human intervention.

  The rack for picking up the containers 1 in the array preferably has the dimensions of a standard microplate, so that the rack is automatically gripped and transported, for example using a microplate handling robot, and a microplate reservoir Alternatively, it is placed in a microplate processing station. A handling system for providing sample aliquots in racks having microplate dimensions is known from patent EP 0 904 841 B1. The rack used therein has a peripheral frame and top and bottom surfaces. The racks comprise grid-like partitions or intermediate walls, which form a plurality of indentations or cavities, each of which can receive a sample tube or container. Special holding means can be removed upwards or downwards and prevents sample tubes that can be inserted from the top or bottom surface from falling out of their cavities.

  Any combination of features of the individual embodiments disclosed herein falls within the scope of the invention. The same reference signs are used for the same features shown in the drawings in each case, even if they do not pay particular attention in the description.

    1 collection / extraction vessel, 2 solid forensic sample, 3 top opening, 4 basically vertical wall, 5 internal chamber, 6 bottom, 7 basket, 8 intermediate floor, 9 upper sample space, 10 lower fluid space, 11 basic Vertical flow path, 12 pipetting needle, 13 top orifice, 14 bottom orifice, 15 partition wall, 16 wall top section, 17 wall bottom section, 18 bottom section top edge, 19 socket, 20 top section Lower end, 21 Fluid collection area, 22 Flat area, 23 Information tag, 24 Closure member, 25 Stopper, 26 Cover, 27 Film, Film structure, 28 Collection set, 29 Collection rack, 30 Compartment, 31 Grasping tube, 32 Extraction set 33 extraction racks 34 intermediate walls 3 5 holding means, 36 upper flange, 37 lower wall thickness region, 38 first protrusion, 39 second protrusion, 40 off-center axis, 41 orientation bar, 42 orientation groove, 43 center axis, 44 shoulder, 50.

Claims (24)

Collect solid forensic sample (2), and / or a container for extracting biological material from their forensic samples (2) (1), this container (1) is, forensic samples ( 2) has an upper opening (3) for insertion, the upper opening (3) being a basically vertical wall (4) surrounding an internal chamber (5) whose depth is limited by the bottom surface (6) The wall (4) and bottom surface (6) are impermeable to fluid, where the container (1) is in contact with the basket (7) in the container (1) and during decomposition / dissolution and extraction. An essentially horizontal intermediate floor (8) for holding a solid forensic sample (2), the intermediate floor (8) being permeable to fluid and upper part of the internal chamber (5) divided into a sample space (9) and the lower fluid space (10), wherein the upper Kiyo device (1) comprises essentially vertical flow path (11), The channel (11) allows the pipetting needle (12) to be inserted through the sample space (9) and into the fluid space (10), and is an upper orifice (3) located near the upper opening (3) of the container (1). 13) and penetrates the intermediate floor (8) at the bottom orifice (14), the vessel (1) also at least partly surrounds the channel (11) from the upper sample space (9) A partition wall (15) separating the latter,
Upper Symbol essentially vertical flow path (11), as the flow path is partially formed by said vertical wall (4) and the partition wall of the container (1) (15), the container Arranged off center with respect to the central axis (43 ) of (1) ,
The basket (7) includes a side wall that is in close contact with the vertical wall (4) or a side wall that is partially formed by the vertical wall, the partition wall (15), and the intermediate floor (8). By forming, the basket (7) has a shape excluding the vertical channel (11),
This allows the basket (7) to be as large as possible for the solid forensic sample (2) introduced into the upper sample space (9) and held by the basket (7).
Container you wherein a. A container (1) for a solid forensic sample (2),
  An essentially vertical wall (4) surrounding the inner chamber (5) and surrounding an upper opening (3) for inserting a solid forensic sample (2);
  The bottom surface (6) limiting the depth of the internal chamber (5), where the bottom surface (6) and the vertical wall (4) are impermeable to fluids,
  An essentially horizontal intermediate floor (8) that is permeable to fluid and divides the internal chamber (5) into an upper sample space (9) and a lower fluid space (10);
  A pipette needle (12) can be inserted into the fluid space (10) through the upper opening (3) of the container (1) and through the sample space (9), and the central axis of the container (1). An essentially vertical access channel (11) arranged off-center with respect to (43);
  Holding the solid forensic sample (2) in the upper sample space (9) and comprising a basket (7) formed by side walls and an intermediate floor (8),
  The vertical access channel (11) is formed by a partition wall (15) and the vertical wall (4), and the partition wall (15) at least partially surrounds the access channel (11), This separates the access channel (11) from the upper sample space (9),
  The side wall of the basket (7) is at least partially surrounded by the partition wall (15) surrounding the access flow path (11) and a wall that adheres most of the wall to the vertical wall (4), Alternatively, the access channel (11) is formed by a wall formed by the vertical wall so that the access channel (11) is introduced into the upper sample space (9) and held by the basket (7). Occupies the minimum space of the sample space (9), allowing the pipetting needle to be inserted through the sample space (9) into the fluid space (10) while making the space for the forensic sample (2) as large as possible;
A container characterized by that. Walls (4) of the container (1), characterized in that it is essentially cylindrical, claim 1 or 2 container according. Basket (7) of the container (1), characterized in that the size to accommodate a buccal swab is determined, container as claimed in any one of claims 1 to 3. Basket (7) is completed as a single injection-molded article of a polymeric material, characterized in that it is inserted into the interior chamber (5) in the container (1), according to any one of claims 1 to 4 container of. The basket (7) is completed as part of the upper section (16) of the wall (4) and is formed with the upper section (16) of the wall (4) as a single injection molded article of polymeric material. wherein, container as claimed in any one of claims 1 to 4. 2. The bottom surface (6) of the container (1) is formed with a wall (4) or a lower section (17) of the wall (4) as a single injection-molded article of polymeric material. container according to any one of 6. The lower section (17) of the wall (4) comprises at its upper end (18) a socket (19) into which the lower end (20) of the upper section (16) of the wall (4) can be sealed. and wherein, container of claim 7 wherein. The bottom surface (6) and / or the lower section (17) of the wall (4) is completed forming a fluid collection area (21) that is the remainder of the bottom surface (6) at the lower level of the container (1), 2. The fluid collection region (21) is present on an essentially vertical axis (40) defined by an essentially vertical channel (11) of the container (1). container according to any of 8. The bottom surface (6) of the container (1) is completed as a film structure (27) glued or welded to the wall (4) or the lower section (17) of the wall (4). container according to any one of 1 to 6. The bottom surface (6) is provided with a flat region (22) on the outside of which the information tag (23) is located, and the information tag (23) includes a 1-D barcode and a 2-D bar code. code, RFID transponder, and characterized in that it is selected from the group comprising RuBee transceiver container according to any one of claims 1 to 10. Forensic samples (2), the gauze pad or buccal swab, findings, filter paper or WHATMAN FTA paper, and characterized in that it is selected from the group comprising textile pieces, according to any of claims 1 to 11 container of. A closure member (24) for the upper opening (3) is provided, the closure member (24) being selected from the group comprising a stopper (25), a cover (26) and a film (27) , container as claimed in any of claims 1 to 12. A collection set having a container (1) according to any of claims 1 to 13 (28), further comprising a collection rack (29) having a compartment (30) for receiving the container (1) A collection set featuring. A extracted set having a container (1) according to any of claims 1 to 13 (32), further comprising a compartment extraction rack having a (30) (33) for receiving the container (1) An extraction set featuring 15. A collection set (28 ) according to claim 14 , wherein the compartment (30) of the collection rack (29 ) is open at the top and bottom, and the container (1) is up or down into the compartment (30). from is insertable, collection set, which is a removable from the compartment downwards or upwards. Collection rack (29), collection set of claim 14 or 16, characterized in that it has the dimensions of standard micro-plate. A method for extracting biological material from a solid forensic sample (2) comprising:
13. (a) Accessable upper opening (3), upper sample space (9) with basket (7), fluid space (10), and flow path (11), according to any of the preceding claims. Providing at least one container (1)
(B) placing a solid forensic sample (2) through the upper opening (3) into the basket (7) in the upper sample space (9) of at least one container;
(C) Disassemble into the basket (7) so that the lower fluid space (10) of the container (1) is filled and the upper sample space (9) is also sufficiently filled to cover the forensic sample (2). / Add lysis buffer;
(D) sealing the entire device (1) and then allowing the culture so that an extraction step is possible;
(E) After culturing, the device (1) is opened or opened, and the amount of degradation buffer solution is taken out by accessing the bottom surface of the lower fluid space (10) through the basically vertical channel (11); (F) Place the device (1) in a centrifuge and rotate so that any degradation / lysis buffer remaining in the solid forensic sample (2) collects at the bottom of the internal chamber (5);
(G) removing the remaining amount of degradation / lysis buffer collected by centrifugation at the bottom of the lower fluid space (10) by accessing it through a basically vertical channel (11);
With
Here the solid forensic sample (2) is held in the basket (7) in the sample space (9) of at least one container (1) during all of steps (c) to (g).
An extraction method characterized by that. (I) The decomposition / lysis buffer solution and the extracted biological material are taken out by the pipetting needle (12) that is inserted into the flow path (11) and reaches the fluid space (10) through the sample space (9). And (ii) performing amplification and / or analysis of the extracted biological material outside the at least one container (1),
The extraction method according to claim 18 , further comprising: 20. Extraction method according to claim 18 or 19 , characterized in that the degradation / lysis buffer is collected down from the solid forensic sample (2) in the fluid collection area (21) at the bottom of the inner chamber (5). Biological material is extracted from the solid forensic sample is a body fluid, cells, DNA, RNA, proteins, characterized in that it is selected microorganisms, and from a group comprising virus, according to any of claims 18 20 Extraction method. The extraction method according to any one of claims 18 to 21 , wherein the extraction method is automatically performed by a robot and a workstation equipped with a corresponding control system. 16. The extraction set (32) according to claim 15, wherein the extraction rack compartment (30) is open at the top and bottom, and the container (1) can be inserted into the compartment (30) from above or below. An extraction set characterized in that it can be taken out downward or upward from this compartment. 24. Extraction set according to claim 15 or 23, wherein the extraction rack (33) has the dimensions of a standard microplate.

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